A large insertion in the Shiga-like toxin 2 gene (stx2) of an Escherichia coli O157:H7 clinical isolate

Karen C. Jinneman, Stephen D. Weagant, Janelle M. Johnson, Sharon L. Abbott, Walter E. Hill, Bradley J. Tenge, Ngoc Lan Dang, Richard Ramsden, Curtis John Omiecinski

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Six clinical Escherichia coli O157:H7 isolates were epidemiologically linked as part of an outbreak in which the most likely source was undercooked ground beef or cross-contamination from the ground beef to other food products at a Mexican-style restaurant. These cultures were analyzed using molecular genetic, immunological and cytotoxicity procedures. All six isolates were confirmed as E. coli O157:H7 and were indistinguishable by pulsed-field gel electrophoresis using XbaI. The results of polymerase chain reaction (PCR) tests, non-isotopic gene probing, reversed passive latex agglutination (RPLA) kit results and Vero cell assays were consistent for the presence of a functional Shiga-like toxin 1 (Stx 1) protein. All six strains produced a stx2 PCR amplicon product; five strains produced a product which was consistent with the predicted amplicon size and one (SEA 6414) produced a much larger PCR product. The SEA 6414 isolate produced a protein reactive with the RPLA kit anti-Stx 2 antibody but was not cytotoxic to Vero cells. Sequencing of this region revealed that this 1310 bp insertion was very similar to a previously identified IS 1203 sequence and the insertion interrupted the carboxyl end of the coding region of the stx2 gene 'A' subunit. Copyright (C) 2000.

Original languageEnglish (US)
Pages (from-to)115-124
Number of pages10
JournalInternational journal of food microbiology
Issue number1-2
Publication statusPublished - Jun 10 2000


All Science Journal Classification (ASJC) codes

  • Food Science
  • Microbiology

Cite this

Jinneman, K. C., Weagant, S. D., Johnson, J. M., Abbott, S. L., Hill, W. E., Tenge, B. J., ... Omiecinski, C. J. (2000). A large insertion in the Shiga-like toxin 2 gene (stx2) of an Escherichia coli O157:H7 clinical isolate. International journal of food microbiology, 57(1-2), 115-124. https://doi.org/10.1016/S0168-1605(00)00237-3